Analysis Of Hair Follicles Research Articles

DosiKit, a New Immunoassay for Fast Radiation Biodosimetry of Hair and Blood Samples

DosiKit is a field radiation biodosimetry immunoassay for fast triage of individuals exposed to external total-body or partial-body irradiation (TBI or PBI). Assay proof-of-concept based on γ-H2AX analysis of human blood samples has been previously described as a promising tool for rapid assessment of TBI. Here, we report on the performance of the assay for PBI based on an analysis of hair follicles irradiated with a 137Cs gamma-ray source, at doses ranging from 0 to 20 Gy and dose rates ranging from ∼0.8 to ∼3 Gy/min. First, we show that the DosiKit protocol allows extraction and analysis of hair follicle proteins. Next, we show that irradiated hair follicles trigger a DNA damage response by inducing dose-dependent γ-H2AX expression. Since γ-H2AX expression strongly decreases 2 to 4 h postirradiation, due to DNA repair, we hypothesized that an antibody targeting the S*/T*Q domains, phosphorylated by ATM for DNA repair activation (pSQTQ), would extend the postirradiation dosimetry time window. DosiKit analysis of pSQTQ in ex vivo irradiated cynomolgus monkey skin explants shows that these sequences are phosphorylated in a dose-dependent manner up to 8 h postirradiation, and that statistically different ranges of external radiation exposure can be distinguished (0-2 Gy, 5-10 Gy, 20 Gy). Since the DosiKit protocol is intended to be used on both blood and hair samples, we also show that SQTQ sequences are phosphorylated dose-dependently in human blood, allowing samples to be classified into three radiation dose ranges (0-0.1 Gy, 0.5-3 Gy and 5-8 Gy). In conclusion, radiation biodosimetry can be performed on both blood and hair samples up to 8 h after exposure using the DosiKit protocol, allowing the concomitant characterization of TBI and PBI for fast and efficient radiological crisis management.

Analysis of the efficiency of hair removal by different optical methods: comparison of Trichoscan, reflectance confocal microscopy, and optical coherence tomography

Noninvasive diagnostic tools, such as Trichoscan®, reflectance confocal microscopy (RCM), and optical coherence tomography (OCT), are efficient methods of hair shaft and growth evaluation. The aim of this study was to carry out a comparative assessment of these three medical procedures by measuring the hair shaft and hair growth after hair removal for a defined period of five days. The application of these techniques was demonstrated by measuring hair growth on the lower leg of six female volunteers. After removal of the hair shaft with a shaving system, the hair follicle infundibula and the length of the growing hairs were measured with the Trichoscan®, RCM, and OCT method. All three methods are reliable hair measuring tools after hair removal. Trichoscan® is best suited in the implementation of hair growth measurement and RCM in the analysis of hair follicles, whereas the OCT system can be consulted as an additional measurement for the evaluation of the hair follicle and length.

Dermatosparaxis in two White Dorper lambs

CASE HISTORY AND CLINICAL FINDINGS: Two White Dorper lambs from the North Island of New Zealand, 2 and 4 weeks of age, were presented with large skin flaps hanging from the flanks, separation of skin from the subcutis over mobile joints, and de-gloving injuries of the limbs and tail. The lambs were subject to euthanasia on humane grounds. PATHOLOGICAL FINDINGS: Large skin tears with associated haemorrhage, periarticular S/C oedema and generalised skin fragility were observed in both lambs at post-mortem examination. Histology of the affected skin revealed diffuse hyalinisation of dermal collagen compared with control lambs, protein-filled peri-adnexal clefts and areas of deep dermal and S/C granulation tissue consistent with previous separation of skin from the subcutis. Analysis of hair follicles, collected from one of the lambs, using a commercially available genetic test in Australia was consistent with the lamb being homozygous for the mutation responsible for ovine dermatosparaxis. DIAGNOSIS: Likely dermatosparaxis. CLINICAL RELEVANCE: These findings strongly suggest that the mutation responsible for dermatosparaxis in White Dorper sheep is present in New Zealand. Dermatosparaxis should be considered when investigating skin fragility in lambs with White Dorper genetics. Confirmation of the disorder is possible through genetic analysis of hair follicles.

Analysis of Hair Follicles in Mutant Laboratory Mice

Hair follicles, or pilosebacous units, are complicated anatomic structures. Analysis of abnormalities of these structures can be difficult without adequate knowledge of normal changes associated with embryonic and postnatal development as well as regular cycling. A variety of systematic approaches to analysis of the pilosebacous unit are provided here.

The role of BMP signalling in the control of ID3 expression in the hair follicle.

Both the production of the hair shaft in anagen and the initiation of a new hair cycle at telogen are the result of reciprocal interactions between the dermal papilla and the overlying epithelial cells. Secreted factors, such as those of the bone morphogenetic protein (BMP) family, play a crucial role in moderating these interactions. Analysis of hair follicles in different stages of the hair cycle showed that BMP signalling was only active during anagen and again during telogen. During catagen, no BMP signalling occurred in the dermal papilla. ID3, a gene expressed in the dermal papilla of both vibrissa and pelage follicles, is a BMP target, and as such, we found that ID3 was expressed from the earliest stages of morphogenesis. During the hair cycle, ID3 was only expressed in the dermal papilla at middle anagen and telogen. To test the significance of ID3 expression in the dermal papilla, we cultured dermal papilla cells and found that ID3 expression fell significantly after a single passage. ID3 expression was returned to in vivo levels in low- and high-passage cells by culturing to high confluence or by the addition of BMP4. These studies reinforce the requirement for active BMP signalling and cell-cell contacts in the dermal papilla during specific stages in the hair cycle.

Quantitative image analysis of hair follicles in alopecia areata.

We took biopsies from similar sites on the scalps of normal controls, normal looking and hair loss areas of patients with alopecia areata. The specimens were sectioned serially and horizontally. We measured morphological parameters, such as the diameter of the hair shaft, the thickness of the inner root sheath, the diameter of the outer root sheath, the irregularity of the outer root sheath and the ratio between the diameters of the hair shaft and the outer root sheath, using a computerized image analysis system. There were significant differences in 5 parameters between hair loss areas and normal controls. The proportions of vellus and telogen hair were significantly higher in the areata areas than in the controls. The 5 morphological parameters and most quantitative-measured parameters of normal-looking areas from patients with alopecia values intermediate between the other groups.

Clinical and biochemical observations on three cases of hypoxanthine-guanine phosphoribosyltransferase deficiency.

Clinical and enzymatic studies on two brothers with severe deficiencies of erythrocyte hypoxanthineguanine phosphoribosyltransferase (HGPRTase) are described, and are compared with similar studies of a classical case of the Lesch-Nyhan syndrome from another family. The two brothers have no neurological abnormalities, only traces of erythrocyte HGPRTase, erythrocyte adenine phosphoribosyltransferase activities approaching the high levels found in the Lesch-Nyhan patient, and similarly raised plasma and urinary concentrations of uric acid. Despite these strong biochemical similarities between the three patients, there were wide differences in the clinical case histories. In both families the enzyme deficiency appeared to be inherited as an X-linked character through asymptomatic carrier females. The relationship of HGPRTase deficiencies to the Lesch-Nyhan syndrome is discussed. Some observations relating to techniques are reported. Cellulose acetate has been found to give much better separations of labelled reaction products in low-level phosphoribosyltransferase assays than filter paper, when used as a supporting medium for electrophoresis. The analysis of hair follicles gives indications of individuals heterozygous for the enzyme deficiency, but the proportion of enzyme-deficient follicles was very small, and the test needs support from studies of other cell types. Using haemolysates, there were signs of a slow indirect conversion of hypoxanthine to inosinic acid, via inosine. Inosine appears to be labelled by a ribosyl-transfer reaction.